金合欢素对肝癌HepG2细胞增殖、凋亡和迁移的影响及机制研究

doi:10.11958/20221115

天津医药 ›› 2023, Vol. 51 ›› Issue (3): 235-239.doi: 10.11958/20221115

金合欢素对肝癌HepG2细胞增殖、凋亡和迁移的影响及机制研究

吴琼(), 李锦源, 黄文涛, 安娜()

广西医科大学附属肿瘤医院药学科（邮编530021）

收稿日期:2022-07-15 修回日期:2022-09-21 出版日期:2023-03-15 发布日期:2023-03-02
通讯作者: 安娜 E-mail:wuqiong6379@163.com;pic1230@qq.com
作者简介:吴琼（1987），女，主治医师，主要从事肿瘤药理学方面研究。E-mail：wuqiong6379@163.com
基金资助:
广西壮族自治区中青年教师基础能力提升项目(KY2016LX037)

Effects of acacetin on proliferation, apoptosis and migration of hepatocellular carcinoma HepG2 cells and its mechanism

WU Qiong(), LI Jinyuan, HUANG Wentao, AN Na()

Department of Pharmacy, Cancer Hospital Affiliated to Guangxi Medical University, Nanning 530021, China

Received:2022-07-15 Revised:2022-09-21 Published:2023-03-15 Online:2023-03-02
Contact: AN Na E-mail:wuqiong6379@163.com;pic1230@qq.com

吴琼, 李锦源, 黄文涛, 安娜. 金合欢素对肝癌HepG2细胞增殖、凋亡和迁移的影响及机制研究[J]. 天津医药, 2023, 51(3): 235-239.

WU Qiong, LI Jinyuan, HUANG Wentao, AN Na. Effects of acacetin on proliferation, apoptosis and migration of hepatocellular carcinoma HepG2 cells and its mechanism[J]. Tianjin Medical Journal, 2023, 51(3): 235-239.

摘要/Abstract

摘要： Objective To investigate the effect of acacetin (Aca) on the proliferation, apoptosis and migration of liver cancer HepG2 cells by regulating PTEN induced kinase 1 (PINK1)/E3 ubiquitin protein ligase (Parkin) pathway mediated mitochondrial autophagy. Methods Liver cancer HepG2 cells were divided into the control group (normally cultured HepG2 cells), the Aca group (10 μmol/L Aca), the PINK1 small interfering RNA negative control (si-NC) group (transfected with si-NC), the PINK1 small interfering RNA (si-PINK1) group (transfected with si-PINK1), the Aca+si-NC group (treated with 10 μmol/L Aca after transfection of si-NC) and Aca+si-PINK1 group (treated with 10 μmol/L Aca after transfection of si-PINK1). Cell counting kit-8 (CCK-8) method was used to detect cell proliferation. Flow cytometry was used to detect cell apoptosis. Transwell test was used to detect cell migration. Transmission electron microscope was used to observe the formation of autophagosomes. Western blot assay was used to detect expression levels of autophagy related proteins [Beclin-1, microtubule associated protein 1 light chain 3 (LC3)-Ⅰ and LC3-Ⅱ] and PINK1/Parkin pathway related proteins in cells. Results Compared with the control group, the survival rate and the number of migrating of HepG2 cells were significantly decreased in the Aca group, and the apoptosis rate, number of autophagy bodies, expression levels of Beclin-1, LC3-Ⅱ/LC3-Ⅰ, PINK1 and Parkin protein were increased significantly (P<0.05). The survival rate and the number of migrating of HepG2 cells were significantly increased in the si-PINK1 group, and the apoptosis rate, number of autophagy bodies, expression levels of Beclin-1, LC3-Ⅱ/LC3-Ⅰ, PINK1 and Parkin protein were decreased significantly (P<0.05). Compared with the Aca group and the Aca+si-NC group, the survival rate and the number of migrating of HepG2 cells were significantly increased in the Aca+si-PINK1 group, and the apoptosis rate, number of autophagy bodies, expression levels of Beclin-1, LC3-Ⅱ/LC3-Ⅰ, PINK1 and Parkin protein were decreased significantly (P<0.05). Conclusion Aca may inhibit the proliferation and migration and promote cell apoptosis of liver cancer HepG2 cells by activating mitochondrial autophagy mediated by PINK1/Parkin pathway.

关键词: 肝肿瘤，实验性, 线粒体，肝, 自噬, 金合欢素, PINK1/Parkin通路, HepG2细胞

Key words: liver neoplasms, experimental, mitochondria, liver, autophagy, acacetin, PINK1/Parkin pathway, HepG2 cells

中图分类号:

R285

图/表 7

表1 各组HepG2细胞存活率比较 (n=6,%,$\bar{x}$±s)

Tab.1 Comparison of the survival rate of HepG2 cells between the six groups

组别	0 h	24 h	48 h
对照组	100.00±0.00	100.00±0.00	100.00±0.00
Aca组	99.98±0.01	51.42±7.05^a	45.19±8.12^a
si-NC组	99.97±0.02	98.91±0.94^b	99.04±0.83^b
si-PINK1组	99.98±0.02	141.63±10.16^abc	157.90±18.44^abc
Aca+si-NC组	99.98±0.03	52.06±7.12^acd	46.03±8.05^acd
Aca+si-PINK1组	99.97±0.01	96.73±3.04^bde	97.86±2.07^bde
F	2.274	195.980^＊＊	133.017^＊＊

表2 各组HepG2细胞凋亡率、迁移细胞数目、自噬小体数量比较 (n=6,%,$\bar{x}$±s)

Tab.2 Comparison of apoptosis rate, number of migrating cells and number of autophagosomes in HepG2 cells between the six groups

组别	细胞凋亡率（%）	迁移细胞数目（个/视野）	自噬小体数量（个/视野）
对照组	13.67±1.25	93.54±7.87	3.27±0.24
Aca组	46.65±3.28^a	34.49±2.85^a	8.79±0.41^a
si-NC组	15.54±1.14^b	95.97±8.12^b	3.41±0.26^b
si-PINK1组	5.42±0.28^abc	124.48±10.34^abc	1.21±0.12^abc
Aca+si-NC组	44.54±3.85^acd	38.84±3.24^acd	8.87±0.40^acd
Aca+si-PINK1组	16.64±1.43^bde	96.50±5.82^bde	4.15±0.27^bde
F	356.201^＊＊	161.704^＊＊	665.042^＊＊

图1 各组HepG2细胞凋亡变化

Fig.1 Apoptosis of HepG2 cells in each group

图2 各组HepG2细胞迁移变化（结晶紫染色，×200）

Fig.2 Migration changes of HepG2 cells in each group (crystal violet staining, ×200)

图3 各组HepG2细胞中自噬小体形成变化（醋酸双氧铀和柠檬酸铅双染色，×10 000）

Fig.3 Changes of autophagosome formation in HepG2 cells in each group (double staining with uranyl acetate and lead citrate, ×10 000)

图4 各组HepG2细胞中Beclin-1、LC3-Ⅰ、LC3-Ⅱ、PINK1、Parkin蛋白表达变化 A：对照组；B：Aca组；C：si-NC组；D：si-PINK1组；E：Aca+si-NC组；F：Aca+si-PINK1组。

Fig.4 Changes of protein expression of Beclin-1, LC3-Ⅰ, LC3-Ⅱ, PINK1 and Parkin in HepG2 cells in each group

表3 Comparison of Beclin-1, LC3-Ⅱ/LC3-Ⅰ, PINK1 and Parkin protein expression levels in HepG2 cells between the six groups

Tab.3 各组HepG2细胞中Beclin-1、LC3-Ⅱ/LC3-Ⅰ、PINK1、Parkin蛋白表达水平比较 (n=6,%,$\bar{x}$±s)

组别	Beclin-1	LC3-Ⅱ/ LC3-Ⅰ	PINK1	Parkin
对照组	0.51±0.08	0.47±0.06	0.35±0.04	0.57±0.07
Aca组	0.91±0.12^a	1.03±0.15^a	0.78±0.09^a	1.09±0.14^a
si-NC组	0.49±0.07^b	0.44±0.06^b	0.33±0.03^b	0.55±0.08^b
si-PINK1组	0.25±0.04^abc	0.16±0.02^abc	0.15±0.01^abc	0.23±0.03^abc
Aca+si-NC组	0.89±0.11^acd	0.99±0.13^acd	0.80±0.09^acd	1.06±0.12^acd
Aca+si-PINK1组	0.53±0.08^bde	0.60±0.08^bde	0.38±0.05^bde	0.54±0.06^bde
F	51.521^＊＊	77.090^＊＊	118.090^＊＊	81.446^＊＊

参考文献 22

[1]	AMINI M, LOOHA M A, ZAREAN E, et al. Global pattern of trends in incidence,mortality, and mortality-to-incidence ratio rates related to liver cancer,1990-2019:a longitudinal analysis based on the global burden of disease study[J]. BMC Public Health, 2022, 2(1):604-629. doi:10.1186/s12889-022-12867-w.
[2]	李彦南, 刘瑞宝. 肝癌介入与免疫联合治疗的机制与进展[J]. 现代肿瘤医学, 2022, 30(14):2658-2661.
	LI Y N, LIU R B. Mechanism and progress of combined intervention and immunotherapy for liver cancer[J]. Journal of Modern Oncology, 2022, 30(14):2658-2661. doi:10.3969/j.issn.1672-4992.2022.14.039.
[3]	RUMGAY H, FERLAY J, MARTEL C, et al. Global,regional and national burden of primary liver cancer by subtype[J]. Eur J Cancer, 2022, 161(1):108-118. doi:10.1016/j.ejca.2021.11.023.
[4]	ZHANG X, XIN Y, YANG Y, et al. Aspartate aminotransferase-to-platelet ratio index for predicting late recurrence of hepatocellular carcinoma after radiofrequency ablation[J]. Int J Hyperthermia, 2022, 39(1):1437-1445. doi:10.1080/02656736.2022.2043457.
[5]	马纳, 李亚静, 范吉平. 金合欢素药理研究进展[J]. 中国现代应用药学, 2018, 35(10):1591-1595.
	MA N, LI Y J, FAN J P. Advances in pharmacological research of acacetin[J]. Chinese Journal of Modern Applied Pharmacy, 2018, 35(10):1591-1595. doi:10.13748/j.cnki.issn1007-7693.2018.10.035.
[6]	ZHANG G, LI Z, DONG J, et al. Acacetin inhibits invasion,migration and TGF-β1-induced EMT of gastric cancer cells through the PI3K/Akt/Snail pathway[J]. BMC Complement Med Ther, 2022, 22(1):10-21. doi:10.1186/s12906-021-03494-w.
[7]	LOU Y, MA C, LIU Z, et al. Antimony exposure promotes bladder tumor cell growth by inhibiting PINK1-Parkin-mediated mitophagy[J]. Ecotoxicol Environ Saf, 2021, 221(1):112420-112429. doi:10.1016/j.ecoenv.2021.112420.
[8]	ZENG W, ZHANG C, CHENG H, et al. Targeting to the non-genomic activity of retinoic acid receptor-gamma by acacetin in hepatocellular carcinoma[J]. Sci Rep, 2017, 7(1):348-360. doi:10.1038/s41598-017-00233-5.
[9]	王婧, 杨雯, 涂琦, 等. 灯盏花素通过调节PINK1/Parkin信号通路介导的线粒体功能诱导人宫颈癌HeLa细胞凋亡[J]. 实用医学杂志, 2020, 36(24):3333-3337.
	WANG J, YANG W, TU Q, et al. Breviscapine induces apoptosis of human cervical cancer HeLa cells by regulating mitochondrial function mediated by PINK1/Parkin signaling pathway[J]. The Journal of Practical Medicine, 2020, 36(24):3333 -3337. doi:10.3969/j.issn.1006-5725.2020.24.006.
[10]	SINGH S, GUPTA P, MEENA A, et al. Acacetin,a flavone with diverse therapeutic potential in cancer, inflammation, infections and other metabolic disorders[J]. Food Chem Toxicol, 2020, 145(1):111708-111725. doi:10.1016/j.fct.2020.111708.
[11]	YUN S, LEE Y J, CHOI J, et al. Acacetin inhibits the growth of STAT3-activated DU145 prostate cancer cells by directly binding to signal transducer and activator of transcription 3 (STAT3)[J]. Molecules, 2021, 26(20):6204-6223. doi:10.3390/molecules26206204.
[12]	PRASAD N, SHARMA J R, YADAV U C S. Induction of growth cessation by acacetin via β-catenin pathway and apoptosis by apoptosis inducing factor activation in colorectal carcinoma cells[J]. Mol Biol Rep, 2020, 47(2):987-1001. doi:10.1007/s11033-019-05191-x.
[13]	JONES A A, GEHLER S. Acacetin and pinostrobin inhibit malignant breast epithelial cell adhesion and focal adhesion formation to attenuate cell migration[J]. Integr Cancer Ther, 2020, 19(1):1-12. doi:10.1177/1534735420918945.
[14]	YAO N, WANG C, HU N, et al. Inhibition of PINK1/Parkin-dependent mitophagy sensitizes multidrug-resistant cancer cells to B5G1,a new betulinic acid analog[J]. Cell Death Dis, 2019, 10(3):232-247. doi:10.1038/s41419-019-1470-z.
[15]	PANIGRAHI D P, PRAHARAJ P, BHOL C S, et al. The emerging, multifaceted role of mitophagy in cancer and cancer therapeutics[J]. Semin Cancer Biol, 2020, 66(1):45-58. doi:10.1016/j.semcancer.2019.07.015.
[16]	XIA J, CHU C, LI W, et al. Mitochondrial protein UCP1 inhibits the malignant behaviors of triple-negative breast cancer through activation of mitophagy and pyroptosis[J]. Int J Biol Sci, 2022, 18(7):2949-2961. doi:10.7150/ijbs.68438.
[17]	张慧蓉, 钱敏, 李萌. 黄芪多糖通过线粒体自噬抑制肝癌肿瘤细胞[J]. 西北药学杂志, 2021, 36(3):426-429.
	ZHANG H R, QIAN M, LI M. Astragalus polysaccharide inhibits liver cancer tumor cells through mitophagy[J]. Northwest Pharmaceutical Journal, 2021, 36(3):426-429. doi:10.3969/j.issn.1004-2407.2021.03.017.
[18]	CHANG S N, KHAN I, KIM C G, et al. Decursinol angelate arrest melanoma cell proliferation by initiating cell death and tumor shrinkage via induction of apoptosis[J]. Int J Mol Sci, 2021, 22(8):4096-4114. doi:10.3390/ijms22084096.
[19]	YOU Y, LI J, CHEN L, et al. Photothermal killing of A549 cells and autophagy induction by bismuth selenide particles[J]. Materials(Basel), 2021, 14(12):3373-3388. doi:10.3390/ma14123373.
[20]	WANG Y, TANG C, CAI J, et al. PINK1/Parkin-mediated mitophagy is activated in cisplatin nephrotoxicity to protect against kidney injury[J]. Cell Death Dis, 2018, 9(11):1113-1126. doi:10.1038/s41419-018-1152-2.
[21]	丁高峰, 郭雷鸣, 柯少瑞, 等. 腮腺多形性腺瘤及癌在多形性腺瘤中PINK1和Parkin的表达及临床意义[J]. 肿瘤防治研究, 2021, 48(5):470-473.
	DING G F, GUO L M, KE S R, et al. Expression and clinical significance of PINK1 and Parkin in pleomorphic adenoma and carcinoma of parotid gland in pleomorphic adenoma[J]. Cancer Research on Prevention and Treatment, 2021, 48(5):470-473. doi:10.3971/j.issn.1000-8578.2021.20.0923.
[22]	D'ONOFRIO N, MARTINO E, MELE L, et al. Colorectal cancer apoptosis induced by dietary δ-valerobetaine involves PINK1/Parkin dependent-mitophagy and SIRT3[J]. Int J Mol Sci, 2021, 22(15):8117-8133. doi:10.3390/ijms22158117.

金合欢素对肝癌HepG2细胞增殖、凋亡和迁移的影响及机制研究

Effects of acacetin on proliferation, apoptosis and migration of hepatocellular carcinoma HepG2 cells and its mechanism

引用本文

RichHTML

PDF (PC)

可视化

摘要/Abstract

使用本文

图/表 7

参考文献 22

相关文章 15

编辑推荐

Metrics

本文评价

[1]	方杰, 黄芮, 郑红慧, 贾倩倩, 鲍静. miR-9-5p靶向TIMP2诱导多发性骨髓瘤细胞自噬和凋亡的机制[J]. 天津医药, 2024, 52(8): 785-790.
[2]	李大强, 李坚, 陆哲明, 曹阳. 毛蕊异黄酮对脊髓损伤大鼠神经元自噬及凋亡的影响[J]. 天津医药, 2024, 52(8): 798-803.
[3]	钟敏, 施震, 周劲松, 李晋杰. GABA信号通路对脓毒症大鼠急性肺损伤内质网应激和线粒体自噬的影响[J]. 天津医药, 2024, 52(7): 733-737.
[4]	陈芋洁, 黄霞, 邓铂林, 贾文文. 金合欢素调节Hippo信号通路对糖尿病视网膜病变大鼠血管生成的影响[J]. 天津医药, 2024, 52(6): 578-583.
[5]	王柯, 叶寒露. 隐丹参酮调节HIF-1α/BNIP3信号通路对兔膝骨关节炎模型软骨细胞自噬和凋亡的影响[J]. 天津医药, 2024, 52(4): 372-378.
[6]	何颖, 张广华, 田立东, 于泳浩. 富氢液通过增加自噬治疗大鼠神经病理性疼痛[J]. 天津医药, 2024, 52(3): 261-265.
[7]	汪爱华, 张飞忠, 王红英. 麝香酮调节SHH介导的自噬对卵巢癌细胞恶性进展的影响[J]. 天津医药, 2024, 52(2): 142-147.
[8]	赵元元, 吴小华. 基于PI3K/Akt/mTOR信号通路探讨LINC00173对多囊卵巢综合征颗粒细胞自噬的影响[J]. 天津医药, 2024, 52(11): 1121-1126.
[9]	张睿, 陈思思, 王彤丹, 于珮. KLF4通过促进自噬减轻高糖浓度下巨噬细胞胆固醇沉积[J]. 天津医药, 2024, 52(10): 1014-1019.
[10]	乔娜, 田英, 陈杨, 郝静. LncRNA MALAT1对PCOS颗粒细胞凋亡、自噬和PI3K/Akt/mTOR通路的影响[J]. 天津医药, 2024, 52(10): 1020-1024.
[11]	廖忠, 廖伟健, 赖国利, 文茵, 苏志威, 曾举浩, 丁洪光. 漆黄素抑制小胶质细胞NLRP3炎症小体活化缓解脓毒症后认知功能损害的研究[J]. 天津医药, 2024, 52(10): 1025-1030.
[12]	杨渊, 宋爽, 陈容, 刘永莲, 刘春燕. 人参皂苷Rg1拮抗亚砷酸钠诱导C57BL/6小鼠肾毒性研究[J]. 天津医药, 2023, 51(8): 820-824.
[13]	黄承军, 徐宇, 秘乐, 王秀军, 刘振峰, 王红嫚. 细胞自噬在急性呼吸窘迫综合征中的研究进展[J]. 天津医药, 2023, 51(6): 668-672.
[14]	王飞, 张小蕾, 李含章, 李亚楠, 胡梦妮, 马骏. 抑制PI3K/Akt/mTOR信号通路对MPP⁺处理的SH-SY5Y细胞自噬、凋亡及PD特征蛋白表达的影响[J]. 天津医药, 2023, 51(5): 449-453.
[15]	耿永芝, 杨利, 李国伟, 张金涛, 程晓磊, 檀立端. 川陈皮素对急性肾损伤大鼠的改善作用及机制研究[J]. 天津医药, 2023, 51(5): 498-503.